期刊
VIROLOGY
卷 408, 期 1, 页码 1-13出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.virol.2010.08.028
关键词
HIV-1; Neutralizing antibody; PBMC assay; HIV reporter virus; Vaccine assessment; Assay standardization; HIV neutralization; Luciferase; Envelope glycoprotein
类别
资金
- NIH Center for HIV/AIDS Vaccine Immunology (CHAVI) [UO1-AI067854]
- Bill AMP
- Melinda Gates Foundation's Collaboration for AIDS Vaccine Discovery (CAVD)/Comprehensive Antibody Vaccine Immune Monitoring Consortium (CAVIMC) [38619]
- UAB Center for AIDS Research [P30-AI-27767]
- UAB Mucosal HIV and Immunobiology Center [R24 DK-64400]
- Department of Veterans Affairs Medical Center, Research Services
- NIH [AI007439]
- Statistical Center for HIV/AIDS Research and Prevention (SCHARP), Fred Hutchinson Cancer Research Center, Seattle, WA
Effective vaccine development for human immunodeficiency virus type 1 (HIV-1) will require assays that ascertain the capacity of vaccine immunogens to elicit neutralizing antibodies (NAb) to diverse HIV-1 strains. To facilitate NAb assessment in peripheral blood mononuclear cell (PBMC)-based assays, we developed an assay-adaptable platform based on a Renilla luciferase (LucR) expressing HIV-1 proviral backbone. LucR was inserted into pNL4-3 DNA, preserving all viral open reading frames. The proviral genome was engineered to facilitate expression of diverse HIV-1 env sequences, allowing analysis in an isogenic background. The resulting Env-IMC-LucR viruses are infectious, and LucR is stably expressed over multiple replications in PBMC. HIV-1 neutralization, targeting TZM-bl cells, was highly correlative comparing virus (LucR) and cell (firefly luciferase) readouts. In PBMC, NAb activity can be analyzed either within a single or multiple cycles of replication. These results represent advancement toward a standardizable PBMC-based neutralization assay for assessing HIV-1 vaccine immunogen efficacy. (C) 2010 Elsevier Inc. All rights reserved.
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